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Human iPSC Modeling Elucidates Mutation-Specific Responses to Gene Therapy in a Genotypically Diverse Dominant Maculopathy

102 Pages Posted: 26 Apr 2019 Sneak Peek Status: Review Complete

See all articles by Divya Sinha

Divya Sinha

University of Wisconsin - Madison - McPherson Eye Research Institute; University of Wisconsin - Madison - Waisman Center

Benjamin Steyer

University of Wisconsin - Madison - McPherson Eye Research Institute; University of Wisconsin - Madison - Wisconsin Institute for Discovery

Pawan K. Shahi

University of Wisconsin - Madison - McPherson Eye Research Institute; University of Wisconsin - Madison - Department of Pediatrics

Katherine Mueller

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Rasa Valiauga

University of Wisconsin - Madison - Waisman Center

Kimberly L. Edwards

University of Wisconsin - Madison - Waisman Center

Cole Bacig

University of Wisconsin - Madison - Waisman Center

Stephanie S. Steltzer

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Sandhya Srinivasan

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Amr Abdeen

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Evan Cory

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Viswesh Periyasamy

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Alireza Fotuhi Siahpirani

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Sushmita Roy

University of Wisconsin - Madison - Wisconsin Institute for Discovery; University of Wisconsin - Madison - Department of Biostatistics

Bikash R. Pattnaik

University of Wisconsin - Madison - McPherson Eye Research Institute; University of Wisconsin - Madison - Department of Pediatrics; University of Wisconsin - Madison - Department of Ophthalmology and Visual Sciences

Krishanu Saha

University of Wisconsin - Madison - McPherson Eye Research Institute; University of Wisconsin - Madison - Wisconsin Institute for Discovery; University of Wisconsin - Madison - Department of Biomedical Engineering

David Gamm

University of Wisconsin - Madison - McPherson Eye Research Institute; University of Wisconsin - Madison - Waisman Center; University of Wisconsin - Madison - Department of Ophthalmology and Visual Sciences

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Abstract

Dominantly inherited disorders are not typically considered therapeutic candidates for gene augmentation (GA). We tested whether GA or genome editing (GE) could serve as a solo therapy for autosomal dominant Best disease (adBD), a macular dystrophy linked to over 100 mutations in the BEST1 gene, which encodes a homo-pentameric calcium-activated chloride channel (CaCC) in the retinal pigment epithelium (RPE). Since no suitable animal models of adBD exist, we generated RPE from patient-derived induced pluripotent stem cells (iPSC-RPE) and found that GA restored CaCC activity and improved rhodopsin degradation in a subset of adBD lines. iPSC-RPE harboring adBD mutations in calcium clasp or chloride binding domains of the channel, but not in a putative structural region, were responsive to GA. However, reversal of the iPSC-RPE CaCC deficit was demonstrated in every adBD line following targeted CRISPR-Cas9 GE of the mutant allele. Importantly, 95% of GE events resulted in premature stop codons within the mutant allele, and single cell profiling demonstrated no adverse perturbation of RPE transcriptional programs post-editing. These results show that GA is a viable approach for a subset of adBD patients depending on the functional role of the mutated residue. Further, GA non-responders are candidates for targeted GE of the mutant allele. Similar scenarios likely exist for other genotypically diverse dominant diseases, expanding the therapeutic landscape for affected patients.

Suggested Citation

Sinha, Divya and Steyer, Benjamin and Shahi, Pawan K. and Mueller, Katherine and Valiauga, Rasa and Edwards, Kimberly L. and Bacig, Cole and Steltzer, Stephanie S. and Srinivasan, Sandhya and Abdeen, Amr and Cory, Evan and Periyasamy, Viswesh and Siahpirani, Alireza Fotuhi and Roy, Sushmita and Pattnaik, Bikash R. and Saha, Krishanu and Gamm, David, Human iPSC Modeling Elucidates Mutation-Specific Responses to Gene Therapy in a Genotypically Diverse Dominant Maculopathy (April 24, 2019). Available at SSRN: https://ssrn.com/abstract=3377258 or http://dx.doi.org/10.2139/ssrn.3377258
This is a paper under consideration at Cell Press and has not been peer-reviewed.

Divya Sinha

University of Wisconsin - Madison - McPherson Eye Research Institute

Madison, WI
United States

University of Wisconsin - Madison - Waisman Center

716 Langdon Street
Madison, WI 53706-1481
United States

Benjamin Steyer

University of Wisconsin - Madison - McPherson Eye Research Institute

Madison, WI
United States

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Madison, WI
United States

Pawan K. Shahi

University of Wisconsin - Madison - McPherson Eye Research Institute

Madison, WI
United States

University of Wisconsin - Madison - Department of Pediatrics

Madison, WI
United States

Katherine Mueller

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Madison, WI
United States

Rasa Valiauga

University of Wisconsin - Madison - Waisman Center

716 Langdon Street
Madison, WI 53706-1481
United States

Kimberly L. Edwards

University of Wisconsin - Madison - Waisman Center

716 Langdon Street
Madison, WI 53706-1481
United States

Cole Bacig

University of Wisconsin - Madison - Waisman Center

716 Langdon Street
Madison, WI 53706-1481
United States

Stephanie S. Steltzer

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Madison, WI
United States

Sandhya Srinivasan

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Madison, WI
United States

Amr Abdeen

University of Wisconsin - Madison - Wisconsin Institute for Discovery ( email )

Madison, WI
United States

Evan Cory

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Madison, WI
United States

Viswesh Periyasamy

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Madison, WI
United States

Alireza Fotuhi Siahpirani

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Madison, WI
United States

Sushmita Roy

University of Wisconsin - Madison - Wisconsin Institute for Discovery

Madison, WI
United States

University of Wisconsin - Madison - Department of Biostatistics

WARF Room 201
610 Walnut Street
Madison, WI 53726
United States

Bikash R. Pattnaik

University of Wisconsin - Madison - McPherson Eye Research Institute ( email )

Madison, WI
United States

University of Wisconsin - Madison - Department of Pediatrics ( email )

Madison, WI
United States

University of Wisconsin - Madison - Department of Ophthalmology and Visual Sciences ( email )

Madison, WI
United States

Krishanu Saha

University of Wisconsin - Madison - McPherson Eye Research Institute ( email )

Madison, WI
United States

University of Wisconsin - Madison - Wisconsin Institute for Discovery ( email )

Madison, WI
United States

University of Wisconsin - Madison - Department of Biomedical Engineering ( email )

1550 Engineering Drive
Madison, WI 53706
United States

David Gamm (Contact Author)

University of Wisconsin - Madison - McPherson Eye Research Institute ( email )

Madison, WI
United States

University of Wisconsin - Madison - Waisman Center ( email )

716 Langdon Street
Madison, WI 53706-1481
United States

University of Wisconsin - Madison - Department of Ophthalmology and Visual Sciences ( email )

Madison, WI
United States

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